Open AccessContinuous microfluidic assortment of interactive ligands (CMAIL)
Author(s) -
Yi-Hsing Hsiao,
Chaoyang Huang,
Chih-Yung Hu,
Yen-Yu Wu,
Chung-Hsiun Herbert Wu,
Chia-Hsien Hsu,
Chihchen Chen
Publication year - 2016
Publication title -
scientific reports
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.24
H-Index - 213
ISSN - 2045-2322
DOI - 10.1038/srep32454
Subject(s) - biopanning , microfluidics , biochip , phage display , antigen , sorting , ligand (biochemistry) , monoclonal antibody , antibody , chemistry , computational biology , chromatography , peptide library , biology , computer science , materials science , nanotechnology , receptor , biochemistry , peptide sequence , genetics , gene , programming language
Finding an interactive ligand-receptor pair is crucial to many applications, including the development of monoclonal antibodies. Biopanning, a commonly used technique for affinity screening, involves a series of washing steps and is lengthy and tedious. Here we present an approach termed continuous microfluidic assortment of interactive ligands, or CMAIL, for the screening and sorting of antigen-binding single-chain variable antibody fragments (scFv) displayed on bacteriophages (phages). Phages carrying native negative charges on their coat proteins were electrophoresed through a hydrogel matrix functionalized with target antigens under two alternating orthogonal electric fields. During the weak horizontal electric field phase, phages were differentially swept laterally depending on their affinity for the antigen, and all phages were electrophoresed down to be collected during the strong vertical electric field phase. Phages of different affinity were spatially separated, allowing the continuous operation. More than 10 5 CFU (colony forming unit) antigen-interacting phages were isolated with ~100% specificity from a phage library containing 3 × 10 9 individual members within 40 minutes of sorting using CMAIL. CMAIL is rapid, sensitive, specific, and does not employ washing, elution or magnetic beads. In conclusion, we have developed an efficient and cost-effective method for isolating and sorting affinity reagents involving phage display.